RT Journal Article
SR Electronic
T1 Robust differentiation of human pluripotent stem cells into endothelial cells via temporal modulation of ETV2 with modified mRNA
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.03.02.973289
DO 10.1101/2020.03.02.973289
A1 Kai Wang
A1 Ruei-Zeng Lin
A1 Xuechong Hong
A1 Alex H. Ng
A1 Chin Nien Lee
A1 Joseph Neumeyer
A1 Gang Wang
A1 Xi Wang
A1 Minglin Ma
A1 William T. Pu
A1 George M. Church
A1 Juan M. Melero-Martin
YR 2020
UL http://biorxiv.org/content/early/2020/03/05/2020.03.02.973289.abstract
AB Human induced pluripotent stem cell (h-iPSC)–derived endothelial cells (h-iECs) have become a valuable tool in regenerative medicine. However, current differentiation protocols remain inefficient and lack reliability. Here, we describe a method for rapid, consistent, and highly efficient generation of h-iECs. The protocol entails the delivery of modified mRNA encoding the transcription factor ETV2 at the intermediate mesodermal stage of differentiation. This approach reproducibly differentiated thirteen diverse h-iPSC lines into h-iECs with exceedingly high efficiency. In contrast, standard differentiation methods that relied on endogenous ETV2 were inefficient and notably inconsistent. Our h-iECs were functionally competent in many respects, including the ability to form perfused vascular networks in vivo. Importantly, timely activation of ETV2 was critical, and bypassing the mesodermal stage produced putative h-iECs with reduced expansion potential and inability to form functional vessels. Our protocol has broad applications and could reliably provide an unlimited number of h-iECs for vascular therapies.